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Comparing Table I. with Table II. it will be seen that the incidence of the Spirochete pertenuis is practically constant in the eruptive elements both in man and in inoculated monkeys. In the monkeys I have experimented with, the eruption does not become general as in man; notwithstanding this, we must admit that in monkeys also, we have to do with a generalized infection as is proved by the presence of the Spirochete pertenuis in the spleen and lymphatic glands.
The Histo-Pathology of Experimental Yaws.
Monkey No. 4 (Mac. pil.).—In this monkey, 19 days after inoculation as already described, a small infiltrated spot appeared at the point of inoculation over the left eyebrow; the lesion became moist, the secretion drying into a thick crust and attaining the size of a sixpenny piece in about two weeks. Two months later, the first lesion being still present of the same size and with the same characters, four more papules appeared, two close to the first lesion and two just above the upper lip. These papules remained always of small dimension and disappeared within three months. It is possible that these four papules were due to auto-inoculation by scratching; it cannot be excluded, however, that they might represent a partial secondary eruption comparable to the general secondary eruption which appears in man; it must be remembered that though the skin lesions in experimental yaws- with the monkeys I have used are generally localized at the point of inocula- tion, the infection is general, as is clearly proved by the presence of the Spirochete pertenuis in the spleen of the animals.
On the 1st June, 1906, the crust from the primary lesion was removed; from the raw elevated granulating surface a piece of tissue was cut, divided into small portions and fixed with various methods (alcohol, sublimate, &c.); then imbedded in paraffin. Sections were stained with various methods (Pappenheim's, &c.). The two papules which appeared above the upper lip were also removed and investigated by the same methods. The results of the histological examination are briefly the following:-
(I.) Primary Lesion.
(a) A well-marked proliferation of the interpapillary processes.
(b) A cellular infiltration consisting of: (1) numerous typical plasma cells, found diffusely with no definite arrangement; (2) some extravasated polymorphonuclear leucocytes; (3) small mononuclear leucocytes, connective tissue cells and a few mast cells. No true giant cells were observed. The fibrous stroma is very delicate and
scarce.
(II.) Papules removed from Lip.
Practically the same result, only the proliferation of the interpapillary pro- cesses is much less marked.
Comparing these results with those found by Macleod, Unna, Nicholls, and myself, in man; it would seem that the histological structure is practically the same in human yaws as well as in experimental yaws.
THE BORNET-GENGOU REACTION IN Yaws.
I have applied to yaws this reaction following the technique used in syphilis by Wassermann, Neisser, and Brück (see Deuts. Med. Woch., 10th May, 1906). As is well known, the principle of the reaction is this: when complement is mixed with the complex antigen + immune body, and afterwards some sensitized red cells are added, no hæmolysis takes place, as the complement has been already taken up by the complex antigen + immune body, and cannot, therefore, get fixed to the hæmolytic receptors.
If the complex antigen + immune body is absent, or only antigen or only immune body is present, then the complement will remain free, and on addition of the sensitized red cells, will get fixed to the hæmolytic receptors and hæmolysis will take place. From the absence or presence of hæmolysis we can, therefore, detect the presence or absence of the complex antigen + immune body. As the following experiments prove, it is possible to demonstrate the existence of specifio yaws anti- bodies and antigen.
Experiment 1.
To the extract of non-ulcerated yaws papules, containing abundantly the Spiro- chate pertenuis, some serum (heated to 55 co.) is added, derived from a monkey
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which had been successfully inoculated with yaws, and which had been afterwards treated at intervals with subcutaneous inoculations of yaws material.
Then some
fresh guinea-pig serum (complement) is added, and, after a certain time, some sensitized red cells, in my case goat red cells, treated with inactivated serum from a rabbit which had been inoculated several times with goat red cells.
Result: No hæmolysis.
The investigation is repeated using the extract of papules taken from six other different cases of yaws.
Result: Constantly the same, viz., no hemolysis.
Experiment II.
Same procedure as in Experiment I. using, instead of the extract of yaws papules, the extract of leprosy nodules.
Result: Well-marked hæmolysis.
Experiment III.
Same procedure, using the extract of nodules taken from a case of pseudo- granuloma pyogenicum.
Result: Hæmolysis.
Experiment IV.
Same procedure, using, instead of the extract of yaws papules, the extract of syphilitic condilomata.
Result: Hæmolysis.
Experiment V.
Same procedure, using the extract of a syphilitic primary sore which presented numerous individuals of Spirochete pallida of Schaudinn.
Result: Hæmolysis.
Experiment VI.
Extract of yaws papules containing the Spirochete pertenuis, + serum (heated to 55 co.) of a monkey immunized for syphilis, + fresh guinea-pig serum, + sensi- tized red cells.
Result: Hæmolysis.
The experiment is repeated, using the extract of papules from six different cases of yaws, always with the same result, viz., hæmolysis. It is to be noted that the serum of the monkey contained with certainty syphilitic antibodies, as by in- activating it and then adding to it the extract of a primary syphilitic sore, then fresh guinea-pig serum (complement), then sensitized red cells, no hæmolysis takes place.
Experiment VII.
Extract of yaws papules, + serum (heated to 55 co.) derived from a normal monkey, fresh guinea-pig serum (complement), + sensitized red cells..
Result: Hæmolysis, well marked.
Experiment VIII.
Extract of spleen juice obtained by puncture of a case of typical yaws, + inactivated serum of a monkey immunized for yaws, complement, + sensitized red cells.
Result: No hæmolysis.
Experiment IX.
Same procedure as in Experiment VIII., using, instead of the serum of a monkey immunized for yaws, the serum of monkeys immunized for syphilis.
Result: Hæmolysis.
The above experiments show that it is possible to detect specific yaws antigen in the yaws papules and in the spleen of cases of yaws; specific yaws antibodies in the blood of monkeys treated with inoculations of yaws material.
The Experiments IV., V., VI., IX. show also that yaws antibodies and antigen are different from syphilis antibodies and antigen, and; therefore, syphilis and yaws cannot be the same entity.